CN106811491A - The Preparation Method And Their Intermediate of optically active beta-hydroxy ester type compound - Google Patents
The Preparation Method And Their Intermediate of optically active beta-hydroxy ester type compound Download PDFInfo
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- CN106811491A CN106811491A CN201510868583.4A CN201510868583A CN106811491A CN 106811491 A CN106811491 A CN 106811491A CN 201510868583 A CN201510868583 A CN 201510868583A CN 106811491 A CN106811491 A CN 106811491A
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- GMAHCICFURREQB-ZDUSSCGKSA-N CC(C)[C@H](CNC(Cc1ccccc1)=O)C(OC)=O Chemical compound CC(C)[C@H](CNC(Cc1ccccc1)=O)C(OC)=O GMAHCICFURREQB-ZDUSSCGKSA-N 0.000 description 1
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- C07C233/46—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom
- C07C233/51—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by carboxyl groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom having the carbon atom of the carboxamide group bound to an acyclic carbon atom of a carbon skeleton containing six-membered aromatic rings
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Abstract
The invention discloses a kind of Preparation Method And Their Intermediate of optically active beta-hydroxy ester type compound.The preparation method is comprised the steps of：In water, in the presence of PA ase, compound (4-1) is hydrolyzed reaction, you can.Preparation method of the invention by the selection to substrate structure, with the specific hydrolase that closes, when the removing of amino protecting group is carried out; methyl in carboxylate methyl ester is not affected, and can directly carry out follow-up cyclization, and step is short; simple to operate, low cost is more suitable for industrialized production.
Description
Technical field
The present invention relates to a kind of Preparation Method And Their Intermediate of optically active beta-hydroxy ester type compound.
Background technology
4-AA (abbreviation 4AA), is a kind of important medical fine chemicals,
It is mainly used in synthesizing the antibiotic of all kinds of training south class, such as Imipenem.Biapenem, Luo Meipeinan and method
La Peinan etc..These medicinal usages are extensive, to Gram-negative bacteria and positive bacteria, aerobic bacteria, anaerobic bacteria
Etc. being respectively provided with broad spectrum high-effect antibacterial action, thus greatly paid attention to by people.At present, in the conjunction of 4-AA
Into in technique, with racemic 2-benzamide methyl -3- carbonyls methyl butyrate (4-AA-1) as raw material
Synthetic route it is comparatively superior, its synthetic route is usually：
In current existing report in method, the removing of amino protecting group is being carried out by compound 4-AA-2
When, the methyl in carboxylate methyl ester is also removed simultaneously, obtains compound 4-AA-3, and it be cyclized instead
When answering prepare compound 4-AA-5, it is still desirable to carboxylic acid is carried out into esterification generation compound 4-AA-4, so
Cyclization again afterwards；Increase which results in step in preparation method, complex operation, cost increases, unfavorable
In industrialized production.
The content of the invention
The technical problems to be solved by the invention are existing with racemic 2-benzamide in order to overcome
Methyl -3- carbonyls methyl butyrate is prepared in the method for 4-AA for raw material, is carrying out ammonia
During the removing of base protection group, the methyl in carboxylate methyl ester is also removed simultaneously so that step increases in preparation method
Many, complex operation, cost increases, and is unfavorable for the defect of industrialized production etc., and provides a kind of optics
The Preparation Method And Their Intermediate of the beta-hydroxy ester type compound of activity.Preparation method of the invention is by right
The selection of substrate structure, with the specific hydrolase that closes, when the removing of amino protecting group is carried out, carboxylic acid
Methyl in methyl esters is not affected, and can directly carry out follow-up cyclization, and step is short, operation
Simply, low cost, is more suitable for industrialized production.
The invention provides a kind of optically active beta-hydroxy ester type compound (2S, 3R) -2- amine methyl -3- ammonia
The preparation method of base methyl butyrate (5-1), it is comprised the steps of：In water, in PA ase
In the presence of, (2S, 3R) -2- (2- phenyl acetamides methyl) -3-hydroxybutyrate methyl esters (4-1) is hydrolyzed
Reaction, you can；
In the present invention, the preparation method of compound (5-1) is preferably comprised the following steps：In water,
PH value be 8.0-9.5 under conditions of, in the presence of PA ase, compound (4-1) is carried out
Described hydrolysis, is obtained compound (5-1)；More preferably comprise the following steps：It is 8.0-9.5 in pH value
Under conditions of, compound (4-1) and the mixed liquor of water mix with PA ase, carry out institute
The hydrolysis stated, is obtained compound (5-1).Wherein, described PA ase is preferably green grass or young crops
Mycin G acylases.Described penicillin G acylase is preferably immobilized penicillin G acylase.
Described immobilized penicillin G acylase can be the conventional immobilization for producing 6-APA in this area
Penicillin G acylase, such as Zhejiang with the wind Hydril Company L. P production for producing 6-APA's
Immobilized penicillin G acylase, or Hu'nan Fulaige Biological Technology Co. Ltd. production for producing
The immobilized penicillin G acylase of 6-APA.The consumption of described water can be not especially limited, as long as not
Influence the carrying out of reaction, you can.The consumption of described PA ase can be the conventional consumption in this area,
It is preferred that described PA ase is 1 with the mass ratio of compound (4-1):25-1:60, more preferably
Ground is 1:30-1:50 (such as 1:40).The temperature of described hydrolysis can be normal for the such reaction in this area
The temperature of rule, preferably 20 DEG C -50 DEG C, be more preferably 30 DEG C -45 DEG C, is most preferably 40 DEG C.Institute
State the process of hydrolysis can be monitored according to the conventional detection method in this area (such as TLC, GC,
HPLC etc.), as the terminal of reaction when typically being disappeared using compound (4-1).Described hydrolysis
Time can be the such reaction in this area conventional time, preferably -24 hours 6 hours, more preferably
It is 8-12 hours, is most preferably 10 hours.
In the preparation method of compound (5-1), the pH value of reaction system is preferably controlled with alkaline matter
Between 8.0-9.5.Described alkaline matter can be the conventional alkaline matter for adjusting pH in this area,
As long as not influenceing reaction is carried out, you can, preferably aqueous dibasic potassium phosphate solution.Described phosphoric acid hydrogen two
The molar concentration and consumption of aqueous solutions of potassium can be not especially limited, as long as the pH of reaction system can be controlled
Value is between 8.0-9.5.The molar concentration of described aqueous dibasic potassium phosphate solution is preferably
0.05mol/L-0.2mol/L (such as 0.1mol/L).Described hydrolysis during carrying out, preferably
Ground, is monitored to the pH value in reaction system, if the pH value of reaction system is less than 8.0, can use ammonia
The pH value of water regulation reaction system is between 8.0-9.5.The concentration and consumption of described ammoniacal liquor can not be made to have
Body is limited, as long as the pH value of reaction system can be adjusted between 8.0-9.5.
After described hydrolysis terminates, the operation of post processing is preferably also can further include.Described
The method and condition of post processing can be the conventional method and condition of this area organic synthesis post processing, the present invention
In, described post processing is preferably comprised the following steps：Reaction solution after hydrolysis is terminated, is carried out
Separation of solid and liquid (such as filter), removes the water (such as vacuum distillation) in filtrate, the residue for obtaining with
Ethanol mixes, stirring, and separation of solid and liquid (for example filtering) is carried out again, removes the ethanol (example in filtrate
Such as vacuum distillation), obtain final product target compound.Wherein, the consumption of described ethanol can be not especially limited,
It is preferred that it is 8mL/g-15mL/g (such as 10mL/g) with the volume mass ratio of compound (4-1).
The preparation method of described (2S, 3R) -2- amine methyl -3- aminobutyric acid methyl esters (5-1), preferably also
Can further include the following steps：In water, under conditions of hydrogen donor and cofactors are present, in carbonyl
In the presence of reductase, 3- oxos -2- (2- phenyl acetamides methyl) methyl butyrate (2-2) be catalyzed also
Original reaction, is obtained described (2S, 3R) -2- (2- phenyl acetamides methyl) -3-hydroxybutyrate methyl esters (4-1)；
In the present invention, the preparation method of compound (4-1) is preferably comprised the following steps：In water,
Under conditions of pH value is 6.0-9.0, under conditions of hydrogen donor and cofactors are present, in carbonyl reduction
In the presence of enzyme, 3- oxos -2- (2- phenyl acetamides methyl) methyl butyrate (2-2) is carried out into described catalysis
Reduction reaction.Wherein, the pH value of described reduction reaction is preferably 7.0-8.5, is more preferably 8.0.
In the preparation method of compound (4-1), described carbonyl reductase is preferably Ke Feier breast bars
The mutant of the carbonyl reductase in bacterium (Lactobacillus kefir) source.Described lactobacillus kefir
The amino acid sequence of the mutant of the carbonyl reductase in source is preferably as application publication number is
Sequence table SEQ NO ID in the Chinese patent application of CN101883846A：6、8、10、12、14、
16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、
50th, shown in 52,54,56,58,60 or 62；More preferably if application publication number is CN101883846A
Chinese patent application in sequence table SEQ NO ID：6th, 8,10,12,14,24,60 or 62 institute
Show, most preferably if application publication number is sequence table SEQ in the Chinese patent application of CN101883846A
NO ID：Shown in 10.Described carbonyl reductase is preferably present in recombinantly expressing the carbonyl reductase
It is in the broken liquid (abbreviation enzyme liquid) of somatic cells after gene engineering colibacillus induced expression, i.e., described
Carbonyl reductase is preferably recombinantly expressing the gene engineering colibacillus induced expression of the carbonyl reductase
The form of the broken liquid of somatic cells afterwards is used.The described genetic engineering for recombinantly expressing the carbonyl reductase
The preparation method of the broken liquid of Escherichia coli induced expression somatic cells can be the conventional method in this area, preferably
Ground comprises the following steps：By the described gene engineering colibacillus for recombinantly expressing carbonyl reductase induction
Zymotic fluid after expression, be centrifuged obtain thalline after mix with water, clasmatosis is carried out using ultrasonic wave,
.Wherein, in the preparation method of the broken liquid of described somatic cells, the consumption of described thalline and water
Relation can be not especially limited, it is preferred that the water is 1mL/g-20mL/g with the volume mass ratio of thalline,
More preferably it is 5mL/g-15mL/g (such as 10mL/g).The described recombination expression carbonyl reductase
The consumption of the broken liquid of somatic cells after gene engineering colibacillus induced expression can be not especially limited, compared with
Goodly, the volume mass ratio of itself and compound (2-2) is 1mL/g-100mL/g, more preferably for
5mL/g-25mL/g, is most preferably 10mL/g.
In the preparation method of compound (4-1), the described gene work for recombinantly expressing the carbonyl reductase
The method and condition of journey Escherichia coli fermentation Fiber differentiation can be the method and condition of this area routine, specifically
Reference can be made to CN101883846A specifications embodiments of page -69 1 and 2 of page 68.
In the preparation method of compound (4-1), the consumption of described water can be not especially limited, preferably
Ground, its volume mass ratio with compound (2-2) is 5mL/g-50mL/g, more preferably for
10mL/g-30mL/g.Described hydrogen donor is preferably isopropanol.The consumption of described hydrogen donor can be
The conventional consumption in this area, it is preferred that described hydrogen donor is preferably 1 with the volume ratio of water:3-1:4.
Under normal circumstances, the consumption of hydrogen donor is more compared with cofactors, therefore, hydrogen donor can be with water together as upper
State the solvent of catalytic reduction reaction.Described cofactors is preferably NaNADP.Described coenzyme because
Son is preferably 0.001g/L-0.1g/L with the mass volume ratio of water, is more preferably 0.005g/L-0.05g/L；
More preferably it is 0.03g/L.The temperature of described catalytic reduction reaction can be the conventional temperature in this area, compared with
It is goodly 20 DEG C -45 DEG C, is more preferably 30 DEG C -40 DEG C, is most preferably 35 DEG C.Described catalysis reduction
The process of reaction can be monitored (such as TLC, GC, HPLC using the conventional detection method in this area
Deng preferably TLC), as the terminal of reaction when typically reacting complete using compound (2-2).Described
The time of catalytic reduction reaction is preferably 4-24 hours, is more preferably 6-12 hours.
In the preparation method of compound (4-1), the pH value of reaction system is preferably controlled with alkaline matter
Between 6.0-9.0.Described alkaline matter can be the conventional alkaline matter for adjusting pH in this area,
As long as not influenceing reaction is carried out, you can, preferably sodium hydrate aqueous solution or ammoniacal liquor.Described hydrogen-oxygen
The molar concentration or consumption for changing sodium water solution or ammoniacal liquor can be not especially limited, as long as reactant can be controlled
The pH value of system is in 6.0-9.0.
In the preparation method of compound (4-1), after described catalytic reduction reaction terminates, preferably also
The operation of post processing can be further included.The method and condition of described post processing can be such anti-for this area
Method and condition that should be conventional, be preferably comprised the following steps：Reaction after catalytic reduction reaction is terminated
Liquid, (preferably carrying out such as more than 70 DEG C of separation of solid and liquid or high temperature using centrifugation makes enzyme to carry out separation of solid and liquid
After denaturation, separation of solid and liquid is carried out using filter paper filtering), filtrate with organic solvent (preferred esters solvent,
Such as ethyl acetate) extraction, after merging organic layer, eaten with saturated sodium bicarbonate aqueous solution, saturation successively
Salt water washing (preferably washed once), after organic layer anhydrous sodium sulfate drying, remove organic solvent (excellent
Choosing removes solvent using vacuum distillation), obtain final product target compound.
In a preferred embodiment of the present invention, in the preparation method of compound (4-1), described catalysis
After reduction reaction terminates, it is preferred that not post-treated, the reaction after directly catalytic reduction reaction is terminated
Liquid, in the presence of PA ase, carries out described hydrolysis, and compound (5-1) is obtained.
The preparation method of described (2S, 3R) -2- amine methyl -3- aminobutyric acid methyl esters (5-1), preferably also
Can further include the following steps：In organic solvent, in the presence of lewis acid, by N- methylols -2-
Phenyl acetamide (1-2) and methyl acetoacetate (2-1) carry out condensation reaction as follows, and institute is obtained
The compound (2-2) stated；
In the preparation method of described compound (2-2), the method and condition of described condensation reaction can
It is the such reaction in this area conventional method and condition, present invention preferably includes the following steps：By chemical combination
Thing (1-2) and the mixed solution of organic solvent, mix with lewis acid and compound (2-1), carry out
Described condensation reaction, you can.Described organic solvent can be the organic molten of the such reaction routine in this area
Agent, preferably halogenated hydrocarbon solvent.Described halogenated hydrocarbon solvent is preferably dichloromethane.It is described
Lewis acid can be the conventional lewis acid of the such reaction in this area, preferably ferric trichloride (FeCl3)。
Described lewis acidic consumption can be the conventional consumption in this area, it is preferred that it is compound (1-2)
The 1%-20% of quality, is more preferably 5%-15%, is most preferably 10%.Described compound (1-2)
With being not especially limited with magnitude relation for described compound (2-1), it is preferred that described chemical combination
The mol ratio of thing (1-2) and described compound (2-1) is 1:1-1:5, more preferably it is 1:1-1:1.5.
The consumption of described organic solvent can be the conventional consumption of the such reaction in this area, it is preferred that itself and chemical combination
The volume mass ratio of thing (1-2) is 1mL/g-50mL/g, is more preferably 5mL/g-30mL/g, most preferably
It is 10mL/g.The temperature of described condensation reaction can be the conventional temperature of the such reaction in this area, preferably
Ground is 10 DEG C -30 DEG C.The process of described condensation reaction can be carried out according to the conventional detection method in this area
Monitoring (such as TLC, GC, HPLC etc.), as reaction when typically being disappeared using compound (2-1)
Terminal.The time of described condensation reaction can be the such reaction in this area conventional time.
In the preparation method of described compound (2-2), after described condensation reaction terminates, preferably
Also can further include the operation of post processing.The method and condition of described post processing can be such for this area
Conventional method and condition are reacted, is preferably comprised the steps of：It is anti-after above-mentioned condensation reaction is terminated
Liquid organic solvent (for example being extracted using esters solvent, such as ethyl acetate) is answered to be extracted, organic layer
Washed (for example wash 2 times), dried (for example dried using anhydrous sodium sulfate or anhydrous magnesium sulfate)
Afterwards, organic solvent (such as vacuum rotary steam removing organic solvent) is removed, you can.
Present invention also offers one kind (2S, 3R) -2- (2- phenyl acetamides methyl) -3-hydroxybutyrate methyl esters (4-1)
Preparation method, it is comprised the steps of：In solvent, under conditions of hydrogen donor and cofactors are present,
In the presence of carbonyl reductase, 3- oxos -2- (2- phenyl acetamides methyl) methyl butyrate (2-2) is carried out
Catalytic reduction reaction as follows, is obtained described (2S, 3R) -2- (2- phenyl acetamides methyl) -3- hydroxyl fourths
Sour methyl esters (4-1)；
Wherein, the method and condition of described catalytic reduction reaction are the same as those described above.
Present invention also offers compound of the one kind as shown in formula (2-2) or the change as shown in formula (4-1)
Compound：
Without prejudice to the field on the basis of common sense, above-mentioned each optimum condition, can be combined, and obtain final product this
Invent each preferred embodiments.
Agents useful for same of the present invention and raw material are commercially available.
Room temperature of the present invention refers to 10-30 DEG C.
In the present invention, the gene engineering colibacillus fermentation inducement culture of the carbonyl reductase is being recombinantly expressed
Method in, the water for using generally deionized water or sterilized water.
Positive effect of the invention is：
Preparation method of the invention, with the specific hydrolase that closes, is being entered by the selection to substrate structure
During the removing of row amino protecting group, the methyl in carboxylate methyl ester is not affected, after can directly carrying out
Continuous cyclization, step is short, simple to operate, and low cost is more suitable for industrialized production.In addition,
Preparation method of the invention, the yield and ee values of target compound is high, by compound (2-2) preparationization
Compound (4-2), the yield of target compound more than 95%, ee values>99.9%.By compound (4-1)
Prepare compound (5-1), more than 95%, two step total recoverys are more than 90% for the yield of target compound.
Specific embodiment
The present invention is further illustrated below by the mode of embodiment, but is not therefore limited the present invention to
Among described scope of embodiments.The experimental technique of unreceipted actual conditions in the following example, according to normal
Rule method and condition, or selected according to catalogue.
Raw material source-information in following embodiments：
Immobilized penicillin G acylase is purchased from Zhejiang Hydril Company L. P with the wind.
The preparation of N- methylol -2- phenyl acetamides (1-2) of embodiment 1
30.0g phenyl acetamides (1-1) are added in 300mL toluene, 8.6g paraformaldehydes are added
With 1.8g sodium acetates, it is warming up to 80 DEG C and reacts 3 hours.Reaction terminates, and is down to and is stirred overnight at room temperature,
There are a large amount of white solids to separate out, filtering, a small amount of toluene of filter cake is washed, 33.0g whites are obtained after vacuum drying
Solid N- methylol -2- phenyl acetamides (1-2), yield 90%.1H NMR(400MHz,DMSO)δ
8.67(s,1H),7.36–7.14(m,5H),5.58(s,1H),4.49(s,2H),3.41(s,2H).；MS
(ESI) m/z=166 (M++1).
3- oxos-the 2- of the embodiment 2 (preparations of (2- phenylacetyls amido) methylbutanoic acid methyl esters (2-2)
10.0g N- methylol -2- phenyl acetamides (1-2) is added in 100ml dichloromethane, is added
1.0g ferric trichlorides and 9.2g methyl acetoacetates (2-1), the mixture room temperature reaction overnight, until former
Material stops reaction after being wholly absent.100mL water and 200mL ethyl acetate are added toward reaction solution, is filled
Divide stirring layering, organic phase is washed with 2 × 50mL, is spin-dried for after anhydrous sodium sulfate drying, obtains 9.4g
Colourless viscous liquid 3- oxos -2- ((2- phenylacetyls amido) methylbutanoic acid methyl esters (2-2), yield 60.0%.1H NMR(400MHz,CDCl3) δ 7.32 (ddd, J=22.5,9.8,4.2Hz, 3H), 7.24-7.18
(m, 2H), 6.16 (s, 1H), 3.83 (t, J=6.3Hz, 1H), 3.67 (s, 3H), 3.64 (dd, J=10.5,
4.3Hz,2H),3.50(s,2H),2.22(s,3H)；MS (ESI) m/z=264 (M++1).
Result under other reaction conditions：
The preparation of embodiment 3 (2S, 3R) -3- hydroxyls -2- ((2- phenylacetyls amido) methyl) methyl butyrate (4-1)
1st, the preparation of enzyme liquid
Culture medium compound method：
LB culture mediums：10g/L tryptones, 5g/L yeast extracts, 10g/L sodium chloride, pH7.2,
121 DEG C of autoclave sterilization 20min；
TB culture mediums：24g/L yeast extracts, 12g/L tryptones, 16.43g/L K2HPO4.3H2O、
2.31g/L KH2PO4, 5g/L glycerine, pH 7.0-7.5,121 DEG C of autoclave sterilization 20min；
Slant medium：10g/L tryptones, 5g/L yeast extracts, 10g/L sodium chloride, 20g/L
Agar powder, eggplant bottle is dispensed into after mixing by 30-40mL liquid amounts, and it is high that setting is positioned over 121 DEG C of high temperature
Pressure sterilizing 20min, adds 100 μ g/mL kanamycin sulfates after cooling, put into inclined-plane, to be condensed
Into solid.
(1) seed activation：Take the genetic engineering large intestine bar for recombinantly expressing carbonyl reductase KRED10
(the Escherichia coli preparation method is this area conventional method to bacterium, expresses carbonyl reductase KRED10
Sequence be specifically shown in application publication number be CN101883846A Chinese patent application in specification the 73rd
The disclosed mutant nucleotide sequence table SEQ ID No.10 from lactobacillus kefir of page table 5) in glycerol stocks pipe
100 μ L culture presevation liquid, on the slant medium in eggplant bottle, are then put with oese uniform application
In 37 DEG C of incubator overnight incubations (18h)；
(2) seed culture：Take 100 mL sterilized waters importing eggplant bottle and make bacteria suspension, take bacteria suspension
50 μ L access the 250mL shaking flasks equipped with 50mL LB culture mediums, 30 DEG C, 220 rpm cultures 16h；
(3) ferment：Above-mentioned cultured seed culture fluid is all accessed equipped with 1L TB culture mediums
5L shaking flasks, after 220rpm cultures 4-6h, add the IPTG (isopropyl-β-D- of 0.3 mM by 37 DEG C
Thiogalactopyranoside) and it is cooled to 28 DEG C, 220rpm Fiber differentiations 12h.
(4) microorganism collection：Zymotic fluid is collected, centrifuge 4000rpm centrifugation 30min are placed in, abandoned
Bacterium mud is collected after clear, -20 DEG C of freezing collections are positioned over.
(5) prepared by enzyme liquid：Thalline 0.5g is weighed, after being subsequently adding the deionized water resuspension of 5mL,
Clasmatosis is carried out by ultrasonic wave, the broken liquid as enzyme liquid for obtaining is positioned over stand-by in ice bath.
2nd, enzyme reaction operation
((2- phenylacetyls amido) methylbutanoic acid methyl esters (2-2) adds 1L shaking flasks to weigh 5.0g 3- oxos -2-
In, add 150mL water to mix, pH8.0 or so is adjusted with 1M sodium hydroxide solutions, it is put into liter in shaking table
Temperature is subsequently adding 50mL isopropanols, 50mL KRED10 enzyme liquids to 35 DEG C, is eventually adding 0.03g/L
NaNADP, that is, start reaction, shaking speed 180rpm, 35 DEG C of temperature.Reaction is passed through after 6 hours
TLC detections are converted completely.Reaction solution is centrifuged off precipitation, water is mutually with 2 × 150mL acetic acid second
Ester is extracted, and the organic phase of merging is washed once with saturated sodium bicarbonate aqueous solution, saturated common salt respectively, nothing
After aqueous sodium persulfate is dried, it is spin-dried for ethyl acetate and obtains 4.8g anhydrous transparency liquid (2S, 3R) -3- hydroxyl -2- ((2-
Phenylacetyl amido) methyl) methyl butyrate (4-1), yield 95%, ee values>99.9%.1H NMR(400MHz,
CDCl3) δ 7.43-7.22 (m, 6H), 6.06 (s, 1H), 3.96 (d, J=3.0Hz, 1H), 3.92-3.79
(m, 2H), 3.61 (s, 3H), 3.60 (s, 2H), 3.32 (ddd, J=14.4,5.1,3.4Hz, 1H), 2.56-
2.38 (m, 1H), 1.20 (d, J=6.3Hz, 3H)；MS (ESI) m/z=266 (M++1).
Wherein, the configuration of compound (4-1), is by the optically-active data and document of compound (5-1)
What (Org.Biomol.Chem., 2007,5,2812-2825) contrast determined.
The preparation of embodiment 4 (2S, 3R) -3- hydroxyls -2- amine methylbutanoic acids methyl esters (5-1)
20.0g (2S, 3R) -3- hydroxyls -2- ((2- phenylacetyls amido) methyl) methyl butyrate (4-1) is added to
In the aqueous dibasic potassium phosphate solution of 0.1M, 0.5g immobilized penicillin G acylases are added, be warming up to 40 DEG C
Reaction 10 hours, pH=8.5 is adjusted in course of reaction with ammoniacal liquor.Reaction terminates, and filtering, filtrate is spin-dried for,
Add 200mL ethanol to stir in residue 30 minutes, be filtered to remove insoluble salt, filtrate is spin-dried for
To target product 10.5g, yield 95.6%.1H NMR(400MHz,CDCl3)δ4.25–4.19(m,
1H), 3.78 (s, 3H), 3.33-3.28 (m, 2H), 2.77-2.73 (m, 1H), 1.28 (d, J=6.3Hz,
3H)；MS (ESI) m/z=148 (M++1),[α]25 D=-6.5 (c 0.8, MeOH)
Result under other reaction conditions：
The preparation of embodiment 5 (2S, 3R) -3- hydroxyls -2- amine methylbutanoic acids methyl esters (5-1)
By 15.0g 3- oxos -2- ((2- phenylacetyls amido) methylbutanoic acid methyl esters (2-2) add 2L shaking flasks in,
Add 500ml water to mix, pH is adjusted to 8.0 or so with 1M sodium hydroxide solutions, be put into liter in shaking table
Temperature is subsequently adding 150mL isopropanols, 150mL KRED10 enzyme liquids to 35 DEG C, is eventually adding 0.03g/L
NaNADP, that is, start reaction, shaking speed 180rpm, 35 DEG C of temperature.Reaction is passed through after 6 hours
TLC detections are converted completely.Be then added to 0.1M aqueous dibasic potassium phosphate solution adjust pH value to
8.0 or so, add 0.3g immobilized penicillin acylated enzyme G, be warming up to 40 DEG C react 10 hours, instead
During answering pH=8.5 is adjusted with ammoniacal liquor.Reaction terminates, and filtering, filtrate is spin-dried for, and is added in residue
150mL ethanol is stirred 30 minutes, is filtered to remove insoluble salt, and filtrate is spin-dried for obtaining target product 7.7g,
Two step yields 91.3%.Hydrogen modal data is shown in embodiment 4.
Comparative example 1 (2S, 3R) -3- hydroxyls -2- ((2-benzamide base) methyl) methyl butyrate (6-2)
Prepare
By 30.0g 3- oxos -2- ((2-benzamide base) methylbutanoic acid methyl esters (6-1) add 3L shaking flasks in,
Add 1000mL water to mix, pH is adjusted to 8.0 or so with 1M sodium hydroxide solutions, be put into liter in shaking table
Temperature is subsequently adding 300mL isopropanols, 300mL KRED10 enzyme liquids to 35 DEG C, is eventually adding 0.03g/L
NaNADP, that is, start reaction, shaking speed 180rpm, 35 DEG C of temperature.Reaction is passed through after 8 hours
TLC detections are converted completely.Reaction solution is centrifuged off precipitation, water is mutually with 2 × 300mL acetic acid second
Ester is extracted, and the organic phase of merging is washed once with saturated sodium bicarbonate aqueous solution, saturated common salt respectively, nothing
After aqueous sodium persulfate is dried, it is spin-dried for ethyl acetate and obtains 25.8g anhydrous transparency liquid (2S, 3R) -3- hydroxyls
- 2- ((2-benzamide base) methyl) methyl butyrate (6-2), yield 85.3%, ee values>99.9%.1H NMR
(400MHz,DMSO-d6)δ8.51(s,1H),7.80–7.78(m,2H),7.54–7.44(m,3H),
5.00(s,1H),3.83–3.81(m,1H),3.63–3.61(m,1H),3.58(s,3H),2.67–2.66
(m, 1H), 1.10 (d, J=6.3Hz, 3H)；MS (ESI) m/z=252 (M++1).
The preparation of comparative example 2 (2S, 3R) -3- hydroxyls -2- amine methylbutanoic acids methyl esters (5-1)
25.8g (2S, 3R) -3- hydroxyls -2- ((2-benzamide base) methyl) methyl butyrate (6-2) is added to
In the aqueous dibasic potassium phosphate solution of 0.1M, 1.0g immobilized penicillin G acylases are added, be warming up to 40 DEG C
Reaction 24 hours, pH=8.5 is adjusted in course of reaction with ammoniacal liquor.TLC determines that reaction can not be carried out,
Reactant (2S, 3R) -3- hydroxyls -2- ((2-benzamide base) methyl) methyl butyrate (6-2) is not changed in, does not have
There is product to generate.
Comparative example 3 (2S, 3R) -2- ((1,3- dioxoisoindole -2- bases) methyl) -3-hydroxybutyrate methyl esters
The preparation of (8-2)
20.0g 2- ((1,3- dioxoisoindole -2- bases) methyl) -3- oxomethyls methyl butyrate (8-1) are added
Enter in 2L shaking flasks, add 800mL water to mix, pH is adjusted to 8.0 or so with 1M sodium hydroxide solutions,
It is put into shaking table and is warming up to 35 DEG C, is subsequently adding 200mL isopropanols, 200mL KRED10 enzyme liquids,
The NaNADP of 0.03g/L is eventually adding, that is, starts reaction, shaking speed 180rpm, 35 DEG C of temperature.
At this moment raw material 2- ((1,3- dioxoisoindole -2- bases) methyl) -3- oxomethyls methyl butyrate (8-1) are found
Solubility is bad in system, and system is muddy, and reaction still has many raw materials anti-after 24 hours
Should.
Claims (11)
1. a kind of optically active beta-hydroxy ester type compound (2S, 3R) -2- amine methyl -3- aminobutyric acid first
The preparation method of ester (5-1), it is characterised in that it is comprised the steps of：In water, in penicillin acyl
Change in the presence of enzyme, (2S, 3R) -2- (2- phenyl acetamides methyl) -3-hydroxybutyrate methyl esters (4-1) is carried out
Hydrolysis, you can；
2. preparation method as claimed in claim 1, it is characterised in that the preparation of compound (5-1)
Method comprises the following steps：In water, under conditions of pH value is 8.0-9.5, in PA ase
Under effect, compound (4-1) is carried out into described hydrolysis, compound (5-1) is obtained；Preferably
Comprise the following steps：Under conditions of pH value is 8.0-9.5, by the mixing of compound (4-1) and water
Liquid, mixes with PA ase, carries out described hydrolysis, and compound (5-1) is obtained.
3. preparation method as claimed in claim 2, it is characterised in that
Described PA ase is penicillin G acylase；Described penicillin G acylase is preferable
Ground is immobilized penicillin G acylase；
And/or, described PA ase and the mass ratio of compound (4-1) is 1:25-1:60, compared with
It is goodly 1:30-1:50；
And/or, the temperature of described hydrolysis is 20 DEG C -50 DEG C, preferably 30 DEG C -45 DEG C；
And/or, the time of described hydrolysis is -24 hours 6 hours, preferably 8-12 hours；
And/or, in the preparation method of described compound (5-1), reaction system is controlled with alkaline matter
PH value between 8.0-9.5；Described alkaline matter is preferably aqueous dibasic potassium phosphate solution；It is described
The molar concentration of aqueous dibasic potassium phosphate solution be preferably 0.05mol/L-0.2mol/L.
4. the preparation method as described in claim any one of 1-3, it is characterised in that described (2S,
3R) preparation method of -2- amine methyl -3- aminobutyric acid methyl esters (5-1) further comprises the following steps：It is molten
In agent, under conditions of hydrogen donor and cofactors are present, in the presence of carbonyl reductase, by 3-
Oxo -2- (2- phenyl acetamides methyl) methyl butyrate (2-2) carries out catalytic reduction reaction as follows, system
Obtain described (2S, 3R) -2- (2- phenyl acetamides methyl) -3-hydroxybutyrate methyl esters (4-1)；
5. preparation method as claimed in claim 4, it is characterised in that described compound (4-1)
Preparation method comprise the following steps：In a solvent, pH value be 6.0-9.0 under conditions of, in hydrogen donor
Under conditions of existing with cofactors, in the presence of carbonyl reductase, by 3- oxos -2- (2- phenylacetyls
Amine methyl) methyl butyrate (2-2) carries out described catalytic reduction reaction.
6. preparation method as claimed in claim 5, it is characterised in that
Described carbonyl reductase be lactobacillus kefir (Lactobacillus kefir) source carbonyl also
The mutant of protoenzyme；The amino acid sequence of the mutant of the carbonyl reductase in described lactobacillus kefir source
Row Ke Feier preferably such as application publication number disclosed in the Chinese patent application of CN101883846A
Sequence table SEQ NO ID in the carbonyl reduction enzyme mutant of lactobacillus source：6、8、10、12、14、16、
18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、50、
52nd, shown in 54,56,58,60 or 62；Described carbonyl reductase is preferably present in recombination expression
In the broken liquid of somatic cells after the gene engineering colibacillus induced expression of the carbonyl reductase；Described
Recombinantly express the broken liquid of the somatic cells after the gene engineering colibacillus induced expression of the carbonyl reductase
Preparation method be preferably comprised the following steps：By the described gene work for recombinantly expressing the carbonyl reductase
Zymotic fluid after journey Escherichia coli induced expression, be centrifuged obtain thalline after mix with water, using ultrasound
Ripple carries out clasmatosis, you can；The described gene engineering colibacillus for recombinantly expressing the carbonyl reductase
The broken liquid of somatic cells after induced expression is preferably with the volume mass ratio of compound (2-2)
1mL/g-100mL/g, is more preferably 5mL/g-25mL/g；
And/or, in the preparation method of compound (4-1), described solvent is the mixing of water and isopropanol
Solvent；In described water and the mixed solvent of isopropanol, the volume ratio of water and isopropanol is preferably
3:1-4:1；
And/or, described hydrogen donor is isopropanol；
And/or, described cofactors is NaNADP；
And/or, described cofactors and the mass volume ratio of solvent is 0.001g/L-0.1g/L, preferably
It is 0.005g/L-0.05g/L；
And/or, the temperature of described catalytic reduction reaction is preferably 20 DEG C -45 DEG C, is more preferably 30 DEG C
-40℃；
And/or, in the preparation method of compound (4-1), the pH of reaction system is controlled with alkaline matter
Value is between 6.0-9.0；Described alkaline matter is preferably sodium hydrate aqueous solution or ammoniacal liquor.
7. preparation method as claimed in claim 5, it is characterised in that the preparation of compound (4-1)
It is not post-treated after described catalytic reduction reaction terminates in method, directly by catalytic reduction reaction knot
Reaction solution after beam, in the presence of PA ase, carries out described hydrolysis, and chemical combination is obtained
Thing (5-1).
8. the preparation method as described in claim any one of 5-7, it is characterised in that described (2S,
3R) preparation method of -2- amine methyl -3- aminobutyric acid methyl esters (5-1) further comprises the following steps：Have
In machine solvent, in the presence of lewis acid, by N- methylol -2- phenyl acetamides (1-2) and acetyl second
Sour methyl esters (2-1) carries out condensation reaction as follows, and described compound (2-2) is obtained；
9. the method for preparing as claimed in claim 8, it is characterised in that described compound (2-2)
Preparation method comprise the following steps：By compound (1-2) and the mixed solution of organic solvent, with road
Lewis acid and compound (2-1) mix, and carry out described condensation reaction, you can；Described is organic molten
Agent is preferably halogenated hydrocarbon solvent；Described halogenated hydrocarbon solvent is preferably dichloromethane；Described
Lewis acid is preferably ferric trichloride；Described lewis acidic consumption is preferably compound (1-2)
The 1%-20% of quality, is more preferably 5%-15%；Described compound (1-2) and described compound
The mol ratio of (2-1) is preferably 1:1-1:5, more preferably it is 1:1-1:1.5；Described organic solvent with
The volume mass of compound (1-2), than preferably 1mL/g-50mL/g, is more preferably 5mL/g-30mL/g；
The temperature of described condensation reaction is preferably 10 DEG C -30 DEG C.
10. the preparation side of one kind (2S, 3R) -2- (2- phenyl acetamides methyl) -3-hydroxybutyrate methyl esters (4-1)
Method, it is comprised the steps of：In solvent, under conditions of hydrogen donor and cofactors are present, in carbonyl
In the presence of reductase, 3- oxos -2- (2- phenyl acetamides methyl) methyl butyrate (2-2) is carried out into following institute
The catalytic reduction reaction for showing, is obtained described (2S, 3R) -2- (2- phenyl acetamides methyl) -3-hydroxybutyrate methyl esters
(4-1)；
Wherein, the condition of described catalytic reduction reaction is as described in claim any one of 4-6.
Compound of 11. one kind as shown in formula (2-2) or the compound as shown in formula (4-1)：
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09143156A (en) * | 1995-11-17 | 1997-06-03 | Tanabe Seiyaku Co Ltd | Production of acetoxyazetidinone derivative and its synthetic intermediate |
| CN1154367A (en) * | 1995-11-17 | 1997-07-16 | 田边制药株式会社 | Process for preparing acetoxyazetidinone derivative and intermediate thereof |
| US20090104671A1 (en) * | 2005-07-20 | 2009-04-23 | Kaneka Corporation | Method for producing optically active 2-(n-substituted aminomethyl)-3-hydroxybutyric acid ester |
| CN103045504A (en) * | 2012-12-05 | 2013-04-17 | 浙江工业大学 | Microorganism catalysis prepared (2S,3R)-2-benzoyl aminomethyl-3-hydroxybutyric acid ester and bacterial strain |
| CN104073543A (en) * | 2014-06-06 | 2014-10-01 | 广东立国制药有限公司 | Method for synthetizing 7-amino-3-vinyl-cephalosporin ring-4-carboxylic acid |
| CN104846025A (en) * | 2015-03-31 | 2015-08-19 | 浙江大学 | Method for preparing (2S, 3R)-2-benzoyl aminomethyl-3-hydroxy methyl butyrate |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU7359198A (en) * | 1997-05-01 | 1998-11-27 | G.D. Searle & Co. | Method and apparatus for preparation of chiral beta amino acids |
| DE10119274A1 (en) * | 2001-04-20 | 2002-10-31 | Juelich Enzyme Products Gmbh | Enzymatic process for the enantioselective reduction of keto compounds |
| JP2004339205A (en) * | 2003-04-24 | 2004-12-02 | Ajinomoto Co Inc | METHOD FOR PRODUCING OPTICALLY ACTIVE N-ARYL-beta-AMINO ACID COMPOUND |
| CN1928102B (en) * | 2006-06-09 | 2011-08-10 | 爱斯特（成都）医药技术有限公司 | Resolution method of beta-amino acid |
| EP2205727B1 (en) * | 2007-10-01 | 2015-06-24 | Codexis, Inc. | Ketoreductase polypeptides for the production of azetidinone |
-
2015
- 2015-12-01 CN CN202210900215.3A patent/CN115057796A/en active Pending
- 2015-12-01 CN CN201510868583.4A patent/CN106811491B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH09143156A (en) * | 1995-11-17 | 1997-06-03 | Tanabe Seiyaku Co Ltd | Production of acetoxyazetidinone derivative and its synthetic intermediate |
| CN1154367A (en) * | 1995-11-17 | 1997-07-16 | 田边制药株式会社 | Process for preparing acetoxyazetidinone derivative and intermediate thereof |
| US20090104671A1 (en) * | 2005-07-20 | 2009-04-23 | Kaneka Corporation | Method for producing optically active 2-(n-substituted aminomethyl)-3-hydroxybutyric acid ester |
| CN103045504A (en) * | 2012-12-05 | 2013-04-17 | 浙江工业大学 | Microorganism catalysis prepared (2S,3R)-2-benzoyl aminomethyl-3-hydroxybutyric acid ester and bacterial strain |
| CN104073543A (en) * | 2014-06-06 | 2014-10-01 | 广东立国制药有限公司 | Method for synthetizing 7-amino-3-vinyl-cephalosporin ring-4-carboxylic acid |
| CN104846025A (en) * | 2015-03-31 | 2015-08-19 | 浙江大学 | Method for preparing (2S, 3R)-2-benzoyl aminomethyl-3-hydroxy methyl butyrate |
Non-Patent Citations (2)
| Title |
|---|
| 孙志浩: "生物催化制备手性化合物技术进展", 《精细与专用化学品》 * |
| 郑裕国等: "手性医药化学品生物催化合成进展与实践", 《生物加工过程》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110093302A (en) * | 2019-06-13 | 2019-08-06 | 浙江华睿生物技术有限公司 | A kind of lactobacillus mutant strain and its application |
| CN110093302B (en) * | 2019-06-13 | 2020-08-28 | 浙江华睿生物技术有限公司 | Lactobacillus mutant strain and application thereof |
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